CN101455310B - Celery extract preparation method and celery extract prepared using the said method - Google Patents
Celery extract preparation method and celery extract prepared using the said method Download PDFInfo
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- CN101455310B CN101455310B CN2009100005343A CN200910000534A CN101455310B CN 101455310 B CN101455310 B CN 101455310B CN 2009100005343 A CN2009100005343 A CN 2009100005343A CN 200910000534 A CN200910000534 A CN 200910000534A CN 101455310 B CN101455310 B CN 101455310B
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Abstract
The invention discloses a celery extract preparation method, including in sequence the following steps: celery beating, heating, compound enzyme enzymolysis, pancreatin enzymolysis, solid-liquid separation, DEAE chromatography column adsorption and hyperfiltration. This extraction method is simple, rapid, safe, non-polluting; and has high yield and low production costs; and the obtained products are nutritious and suitable for large-scale industrial production.
Description
Technical field
The present invention relates to the production technology of food, biological medicine, specifically the preparation method of Herba Apii graveolentis extract.
Background technology
Celery has the branch of Chinese celery and celery, is samphire.It also is the daily delicacies on our dining table.It not only can be cooked, but hyoscine again, and medicinal based on celery morning, celery fragrance is denseer, also claims " Sheep's-parsley ".Because it is medicinal that their root, stem, leaf and seed all can be worked as, so the title of " medicine in the kitchen ", " medicine celery " is arranged again.Really, the pharmacological action of celery is very wide, and higher use value is arranged.
The nutritional labeling that celery contains is very important to human body, active material wherein also is to guarantee that the healthy institute of human body is indispensable, its pharmacological action is the result of multiple composition effect, for example the Flavonoid substances in the celery, butyl benzene phthaleins, amino acid, unrighted acid isoreactivity material all have the physiological function of multiple pharmacologically actives such as antibiotic, anti-oxidant, antitumor, reducing blood lipid, so the hypertension and hyperlipemia curative effect that Herba Apii graveolentis extract has is the coefficient result of multicomponent.
Summary of the invention
The objective of the invention is to propose a kind of from the celery raw material method of separation and Extraction Herba Apii graveolentis extract, adopt enzymolysis and chromatography method, as much as possible flavone compound useful in the celery, small-molecular peptides, active polysaccharide class, trace element, amino acid etc. are extracted.
Extracting method of the present invention comprises following process successively:
(1) get celery, clean a certain amount of water making beating of adding, slurries are heated to 85 ℃～100 ℃, and be more than the 10min, to be cooled to room temperature heat time heating time, and slurries are standby;
(2) slurry temperature remains on 30 ℃～55 ℃, transfers pH to 3.0～5.5 with acid, adds the complex enzyme that contains cellulase, hemicellulase and pectase, after stirring evenly, keeps pH to 3.0～5.5, enzymolysis under 30 ℃～55 ℃ condition;
(3) transfer pH to 8.0～9.5 with the slurries of alkali after with enzymolysis, add pancreatin, after stirring evenly, keep the condition enzymolysis of pH to 8.0～9.5,45 ℃～55 ℃, enzymolysis finishes the back heat temperature raising and carries out enzyme-deactivating, is cooled to room temperature;
(4) with pH to 6.0～7.2 of enzymolysis liquid behind the enzyme-deactivating in the sour pacing rapid (3), carry out Separation of Solid and Liquid, remove the celery residue, collect filtered solution, standby;
(5) filtered solution is adsorbed with the DEAE chromatographic column, use the superphosphate buffer solution elution;
(6) eluent carries out ultrafiltration with filter membrane, collects filter liquor, promptly gets the Herba Apii graveolentis extract concentrate.
The part by weight of cellulase, hemicellulase and pectase is 1～3: 1～3 in the described complex enzyme: 1～2, be preferably 2: 2: 1, can also contain amylase in the complex enzyme, complex enzyme makes the nutritional labeling of celery be extracted out fully, make the utilization rate of celery higher, comprehensively keep the nutritional labeling of celery.
The alkali that described adjust pH is used can be used NaOH, calcium hydroxide, potassium hydroxide, sodium acid carbonate or ammoniacal liquor.
The acid that described adjust pH is used can be used hydrochloric acid, phosphoric acid, citric acid or sulfuric acid.
Carry out the process of enzymolysis with enzyme, can grasp the enzymolysis process by the control enzymolysis time, enzymolysis time is short, and the insufficient yield that causes of enzymolysis is low, but the enzymolysis time process can extend manufacture cycle again, increases cost and increases contaminated chance.Enzymolysis time was advisable with 60～210 minutes in the described step (2).Enzymolysis time was advisable with 150～240 minutes in the described step (3).The enzyme-deactivating temperature is 90 ℃～100 ℃ in the described step (3), and inactivation time is 5～15 minutes.
Separation of Solid and Liquid in the described step (4) can be that first suction filtration gets supernatant, and residue is centrifugal with centrifuge, collects supernatant, merges the supernatant of suction filtration and centrifugal gained, filters, and isolates clear liquid.
Described acid phosphatase salt buffer is preferably pH6.0～6.8 phosphate buffers.
The preferred molecular cut off of described ultrafiltration is the milipore filter of 50～100KD.
Concentrate can be used as the direct can of liquid extract, perhaps further dry the processing.
The present invention also provides a kind of and has extracted the Herba Apii graveolentis extract that obtains by said method, is to contain flavone compound, small-molecular peptides, active polysaccharide class, trace element, amino acid isoreactivity material.
The present invention has following advantage:
1, owing to content of cellulose in the celery is higher, celery of the present invention is directly through two step enzymolysis and extraction Herba Apii graveolentis extracts, the first step is carried out enzymolysis with the complex enzyme of cellulase, hemicellulase and pectase earlier, can be with cellulose, the pectin enzymolysis of celery, the water-soluble fibre of celery and flavone compound etc. are fully extracted.Second step was adopted the pancreatin enzymolysis, can obtain small-molecular peptides and micromolecular polysaccharide.
2, celery homogenate behind the two step enzymolysis, adopt extraction, the centrifugal acquisition supernatant of residue, after filtration, go up EDAE chromatographic column absorption and purification, production technologies such as ultrafiltration, enzymolysis is complete, can remove big molecular impurity, obtain nutritious extract, wherein contain flavone compound, small-molecular peptides, active polysaccharide class, trace element, amino acid isoreactivity material.
3, adopt two step enzymolysis, extract, the centrifugal acquisition supernatant of residue, after filtration, go up DEAE chromatographic column absorption and purification, production technologies such as ultrafiltration, advantage is that yield height, production cost are low, production environment requires lowly, has reduced product and has been subjected to microbial contamination, rotten chance.
4, celery directly extracts Herba Apii graveolentis extract through production technologies such as two step enzymolysis, and the nutriment with celery extracts to greatest extent.Mainly contain flavone compound, small-molecular peptides, active polysaccharide class, trace element, amino acid isoreactivity material in the extract that obtains, the medical value height, nutriment is abundanter, easier is absorbed by human body, is easilier accepted by market and consumer.
The specific embodiment
Below by specific embodiment the present invention is described in further detail.
Embodiment:
Get the celery of 2.0kg, clean, add the water making beating of 18.0kg, weigh 20.0kg.Slurries are heated to 85 ℃～100 ℃, preferably are controlled to be 95 ℃, keep 15min.Be cooled to 50 ℃, slurries are standby.
Transfer pH3.0～5.5 with 4.5% (w%) hydrochloric acid, preferably transfer to 3.8 ± 0.2, the ratio that adds 10 million international units (than vigor) complex enzyme in every kg celery, add 20 million international units (than vigor) complex enzyme, the part by weight of cellulase, hemicellulase and pectase is 2: 2: 1 in the complex enzyme, after stirring evenly, keep pH 3.8 ± 0.2 keeping its stabilized enzyme to separate environment, enzymolysis 90min under 50 ℃ ± 0.5 ℃ the condition.Hydrolysis temperature can be controlled under 30 ℃～55 ℃ conditions, and enzymolysis time can suitably be adjusted in this scope, and the high time of temperature can suitably shorten, and temperature is hanged down the time proper extension.
With concentration is that the NaOH of the 18g/ml slurries after with enzymolysis are transferred pH to 8.0～9.5, preferably control pH to 9.0 ± 0.2, add the ratio of 5 million international units (than vigor) pancreatin in every kg celery, add 10 million international units (than vigor) pancreatin, after stirring evenly, keep pH to 9.0 ± 0.2,45 ℃～55 ℃ condition enzymolysis preferably is controlled at 50 ℃ ± 0.5 ℃ condition enzymolysis, and enzymolysis finishes back heat temperature raising to 100 ℃, be incubated 10 minutes and carry out enzyme-deactivating, be cooled to room temperature.
Transfer pH to 6.0～7.2 with 4.5% (w%) hydrochloric acid, preferably control pH to 6.5 ± 0.2, suction filtration gets supernatant 14.0kg, and residue is centrifugal with centrifuge, collects supernatant and gets 3.0kg, merges supernatant, altogether 17.0kg.Carry out coarse filtration, collect filter liquor and get 16.0kg.
With DEAE furnishing pasty state, adopt slurry type method dress post with water for injection, dress column pressure 0.5Mpa.Clean with 400mmol/L pH6.5 phosphate buffer earlier behind the dress post, use 20mmol/L pH6.5 phosphate buffer balance again, DEAE chromatographic column on the above-mentioned filter liquor is adsorbed, with 20mmol/L pH6.5 phosphate buffer wash-out.Collect object and get 15.0kg, use the filter membrane that molecule is held back to 100K to carry out ultrafiltration, collect filter liquor and get 13.0kg, promptly get the Herba Apii graveolentis extract concentrate, sampling detects.The usually phosphate buffer solution can be selected pH6.0～6.8.
Assay:
1. polysaccharide is differentiated: phenolsulfuric acid reagent reacting (positive)
2. polypeptide is differentiated: get this product 1ml, add ninhydrin solution number droplet, heating shows purple, illustrates that this product is for containing polypeptide solution.
3.pH value is 6.5 (two appendix VI of Chinese Pharmacopoeia version in 2005 H)
4. flavonoid content is measured: (with the rutin is that reference substance is measured content of total flavone in the Herba Apii graveolentis extract to 0.28mg/ml, add aluminium ion reagent, control the appropriate pH value simultaneously, make chromocor compound and aluminium salt form complex compound, can obtain the stable characteristics absworption peak at visible region.)
5. micro-: Zn:0.1083mg/ml; Fe:0.0112mg/ml; Mn:0.0037mg/ml; Cu:0.0021mg/ml.(atomic absorption spectrography (AAS))
Claims (9)
1. the preparation method of a Herba Apii graveolentis extract is characterized in that this method comprises following process successively:
(1) get celery and clean, add a certain amount of water making beating, slurries are heated to 85 ℃～100 ℃, and be more than the 10min, to be cooled to room temperature heat time heating time, and slurries are standby;
(2) slurry temperature remains on 30 ℃～55 ℃, transfers pH to 3.0～5.5 with acid, adds the complex enzyme that contains cellulase, hemicellulase and pectase, after stirring evenly, keep pH to 3.0～5.5, enzymolysis under 30 ℃～55 ℃ condition, enzymolysis time are 60～210 minutes;
(3) transfer pH to 8.0～9.5 with the slurries of alkali after with enzymolysis, add pancreatin, after stirring evenly, keep the condition enzymolysis of pH to 8.0～9.5,45 ℃～55 ℃, enzymolysis time is 150～240 minutes, and enzymolysis finishes the back heat temperature raising and carries out enzyme-deactivating, is cooled to room temperature;
(4) with pH to 6.0～7.2 of enzymolysis liquid behind rapid (3) enzyme-deactivating of sour pacing, carry out Separation of Solid and Liquid, remove the celery residue, collect filtered solution, standby;
(5) filtered solution is adsorbed with the DEAE chromatographic column, use the superphosphate buffer solution elution;
(6) eluent carries out ultrafiltration with filter membrane, collects the filter liquor that obtains after the ultrafiltration, promptly gets the Herba Apii graveolentis extract concentrate.
2. preparation method as claimed in claim 1 is characterized in that: the part by weight of cellulase, hemicellulase and pectase is 1～3: 1～3 in the complex enzyme in the described step (2): 1～2.
3. preparation method as claimed in claim 1 or 2 is characterized in that: the alkali that described adjust pH is used is NaOH, calcium hydroxide, potassium hydroxide, sodium acid carbonate or ammoniacal liquor.
4. preparation method as claimed in claim 1 or 2 is characterized in that: the acid that described adjust pH is used is hydrochloric acid, phosphoric acid, citric acid or sulfuric acid.
5. preparation method as claimed in claim 1 or 2 is characterized in that: the enzyme-deactivating temperature is 90 ℃～100 ℃ in the described step (3), and inactivation time is 5～15 minutes.
6. preparation method as claimed in claim 1 or 2 is characterized in that: the Separation of Solid and Liquid in the described step (4) is that first suction filtration gets supernatant, and residue is centrifugal with centrifuge, collect supernatant, merge the supernatant of suction filtration and centrifugal gained, filter, isolate clear liquid.
7. preparation method as claimed in claim 1 or 2 is characterized in that: described acid phosphatase salt buffer is elected pH6.0～6.8 phosphate buffers as.
8. preparation method as claimed in claim 1 or 2 is characterized in that: it is the milipore filter of 50～100KD that molecular cut off is selected in described ultrafiltration.
9. the Herba Apii graveolentis extract of any described method preparation in the claim 1 to 8.
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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101978979B (en) * | 2010-11-03 | 2012-01-04 | 隋焕平 | Method for preparing refined celery extract |
| CN102318846B (en) * | 2011-06-07 | 2013-04-17 | 韩金光 | Processing method of jack-fruit |
| CN105028811A (en) * | 2015-07-06 | 2015-11-11 | 北京京隆卓尚投资有限公司 | Herba cistanche tartary buckwheat tea, and preparation method and applications thereof |
| CN105595128A (en) * | 2016-01-14 | 2016-05-25 | 山东鲁菱果汁有限公司 | Concentrated clear celery juice and preparation method thereof |
| CN105919023A (en) * | 2016-05-03 | 2016-09-07 | 王胜 | Plant salt containing celery extract, and preparation method of plant salt |
| CN105942433A (en) * | 2016-05-03 | 2016-09-21 | 王胜 | Plant salt containing pine pollen extracts and preparation method thereof |
| CN106389489A (en) * | 2016-09-20 | 2017-02-15 | 青海清华博众生物技术有限公司 | Celery-seed extract and extracting method and application thereof |
| CN107080256A (en) * | 2017-05-17 | 2017-08-22 | 北京泓九生物科技有限公司 | A kind of compound of walnut oligopeptide and date polysaccharide and preparation method thereof |
| CN108842297B (en) * | 2018-07-20 | 2020-02-21 | 哈工大机器人（山东）智能装备研究院 | Celery fiber mask base cloth and preparation method thereof |
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| CN1250612A (en) * | 1999-04-21 | 2000-04-19 | 吴坤 | Process for extracting active components from celery for further processing |
| CN1392146A (en) * | 2002-06-11 | 2003-01-22 | 张在雪 | Water cress extract and its preparing method and use |
| CN101228942A (en) * | 2008-02-21 | 2008-07-30 | 伍曾利 | Method of preparing banana extract |
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Patent Citations (3)
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|---|---|---|---|---|
| CN1250612A (en) * | 1999-04-21 | 2000-04-19 | 吴坤 | Process for extracting active components from celery for further processing |
| CN1392146A (en) * | 2002-06-11 | 2003-01-22 | 张在雪 | Water cress extract and its preparing method and use |
| CN101228942A (en) * | 2008-02-21 | 2008-07-30 | 伍曾利 | Method of preparing banana extract |
Non-Patent Citations (2)
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| JP特开2003-135000A 2003.05.13 |
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